Studies using an in vitro model show evidence of involvement of epithelial-mesenchymal transition of human endometrial epithelial cells in human embryo implantation.

Human embryo implantation is a critical multistep process consisting of embryo apposition/adhesion, followed by penetration and invasion. Through embryo penetration, the endometrial epithelial cell barrier is disrupted and remodeled by an unknown mechanism. We have previously developed an in vitro model for human embryo implantation employing the human choriocarcinoma cell line JAR and the human endometrial adenocarcinoma cell line Ishikawa. Using this model we have shown that stimulation with ovarian steroid hormones (17ß-estradiol and progesterone, E2P4) and suberoylanilide hydroxamic acid (SAHA), a histone deacetylase inhibitor, enhances the attachment and adhesion of JAR spheroids to Ishikawa. In the present study we showed that the attachment and adhesion of JAR spheroids and treatment with E2P4 or SAHA individually induce the epithelial-mesenchymal transition (EMT) in Ishikawa cells. This was evident by up-regulation of N-cadherin and vimentin, a mesenchymal cell marker, and concomitant down-regulation of E-cadherin in Ishikawa cells. Stimulation with E2P4 or SAHA accelerated Ishikawa cell motility, increased JAR spheroid outgrowth, and enhanced the unique redistribution of N-cadherin, which was most prominent in proximity to the adhered spheroids. Moreover, an N-cadherin functional blocking antibody attenuated all events but not JAR spheroid adhesion. These results collectively provide evidence suggesting that E2P4- and implanting embryo-induced EMT of endometrial epithelial cells may play a pivotal role in the subsequent processes of human embryo implantation with functional control of N-cadherin.

Evaluation of a community intervention program in Japan using Framingham risk score and estimated 10-year coronary heart disease risk as outcome variables: a non-randomized controlled trial.

BACKGROUND: Community-based programs are being widely adopted in the struggle to prevent cardiovascular diseases. No study has been conducted in Japan to evaluate the effects of a community-based health promotion program by using the Framingham risk score and 10-year CHD risk as outcome variables. The aim of the present study was to assess the effects of a program involving 6-month intervention and 18-month follow-up using such outcomes. METHODS: Participants (n = 1,983, 39.5% women, mean age 63.4 years) were selected for the study in 2008. Of these 1,983, 347 (42.4% women) subjects received the 6-month intervention. The intervention included individual counseling and group sessions, among others. After 18 months, 1,278 participants (intervention group: 238, control group: 1,040) were followed up. Changes in the Framingham risk score and 10-year coronary heart disease (CHD) risk were evaluated. ANCOVA and multiple logistic models adjusted for baseline value, age, sex and intervention times were used. RESULTS: The results showed that the differences in the Framingham risk score and mean 10-year CHD risk were significant in the intervention group compared with the control group after 6-month follow-up (-0.46 and -1.12, respectively) and were also significant after 18-month follow-up (-0.39 and -0.85, respectively). The proportion of those with intermediate 10-year CHD risk (> = 10%) was significantly lower at 6 months (OR 0.30, 95% CI 0.12-0.74) and at 18 months (OR 0.41, 95% CI 0.19-0.92). CONCLUSIONS: The six-month intervention program effectively decreased estimated 10-year CHD risk and the effects were still present at 18-month follow-up. TRIAL REGISTRATION: UMIN-CTR: UMIN000008163.

[Changes in measurements related to metabolic syndrome among individuals with national health insurance after specific health guidance].

OBJECTIVES: In 2008, "Specific health checkup and guidance," a policy that focused on improving the status of metabolic syndrome (MetS), was launched in Japan. The aim of the present study was to evaluate the effects of a 6-month intervention based on this policy on the participants' metabolic syndrome, obesity risk factors, and lifestyles. METHODS: A quasi-experimental design was used between 2008 and 2009. In total, 500 subjects and 1,483 residents of Soka city, Saitama Prefecture were judged to require an active and motivational support (ACS and MOS, respectively) based on a specific health guidance standard in Japan. Of these residents, 72 and 275 individuals attended a program that included individual counseling and a 6-month intervention, respectively. Those who did not attend any intervention and received only information concerning MetS were classified as the control group. Changes in the risk factors related to obesity and MetS were analyzed using the analysis of covariance and multiple logistic models. RESULTS: Of the 347 participants, 62 (86.1%) and 41 (56.900) receiving ACS and 266 (96.700) and 210 (76.400) receiving MOS finished the 6-month intervention program and subsequent follow-up, respectively. After the 6-month intervention, improvement in obesity-related risk factors with ACS was significantly greater than that with MOS. Compared to the control group, those receiving ACS demonstrated significant improvements in the waist circumference (-3.1 cm, P < 0.001), body mass index (BMI; -0.8 kg/m2, P < 0.001), weight (-2.3 kg, P < 0.001), and HbA1c levels (-0.18%, P = 0.016). Those receiving MOS showed significant improvements in the waist circumference (-1.3 cm, P = 0/001), BMI (-0.5 kg/m2, P < 0.001), weight (-1.2 kg, P < 0.001), systolic and diastolic blood pressure (-2.4 mmHg, P = 0.018; -1.8 mmHg, P = 0.005), and HbA1c levels (-0.06%, P = 0.025) after adjusting for sex, age, and baseline values. The proportion of participants receiving ACS and MOS who met the definition of Mets or pre-Mets at the baseline, but did not achieve this threshold after 1 year, was significantly higher than those participants in the control group. The odds ratio with 9500 confidence intervals was 1.41 (1.05-1.90) for combined ACS and MOS, and 1.39 (1.00-1.94) for MOS, respectively. Conclusion Thus, our program helped in improving factors associated with MetS.

Community-based lifestyle modification of cardiovascular disease risks in middle-aged Japanese: a 27-month update.

Lifestyle modification is the cornerstone of preventive management for people with cardiovascular disease risks, such as obesity, hypertension, dyslipidemia and diabetes. This study investigated the effect of a 27-month community-based lifestyle intervention on the reduction of cardiovascular disease risks in middle-aged Japanese. Of 549 participants with cardiovascular disease risk factors of overweight, hypertension, dyslipidemia or diabetes enrolled in this non-randomized controlled study, 397 participants aged 39-71 years old completed all 3 serial surveys at baseline, 15 months and 27 months. For the intervention group (39 males and 174 females), 31 specific interventions including individual counselling and group sessions were conducted. The control group (64 males and 120 females) only received 7 newsletters providing health information and results of health checkups. Multiple logistic regression analysis adjusted for sex, each baseline risk category and age category showed that the proportion of those who were overweight or showed dyslipidemia risk were significantly lower in the intervention group only at 27 months [Odds ratio (OR): 0.43 (95% CI 0.20-0.94), OR: 0.43 (95% CI 0.21-0.87), respectively] and the proportion of those showing diabetes risk was significantly lower in the intervention group at both 15 months [OR: 0.42 (95% CI 0.18-0.97)] and 27 months [OR: 0.56 (95% CI 0.32-0.99)]. In conclusion, the 27-month community-based lifestyle modification of cardiovascular disease risks shows significant reductions in risks of diabetes, overweight and dyslipidemia in middle-aged Japanese.

Fifteen-month lifestyle intervention program to improve cardiovascular risk factors in a community population in Japan.

Morbidity and mortality associated with cardiovascular disease remain primary public health problems in Japan. We developed a large-scale community-based 15-month intervention program, and evaluated its long-term effects on improving lifestyle and cardiovascular risk. Of 549 participants, 436 (46 males and 186 females in intervention group and 72 males and 132 females in control group) completed the study. The intervention program consisted of a 6-month program including 15 sessions, with one lecture and individual counseling, two workshops and nutrition educations, and nine exercises (total 29 hours) and a 9-month maintenance program including 4 sessions of individual counseling, workshop, nutrition, and exercise (total 7 hours). Changes from the baseline to 15 months were significantly different between intervention and control groups by gender: the proportion of subjects who walked more than 6,000 steps/day (+39% vs. +2% for males; +17% vs. +1% for females), exercised more than 20 min/day and 3 times/week (+22% vs. +1% for males; +22% vs. +8% for females), ate vegetables at every meal (+18% vs. +4% for females), and consumed less salt (+9% vs. +3% for females); and the changes in body weight (-1.0 vs. -0.1 kg for males; -1.8 vs. -0.6 kg for females). Other changes detected only in females were body mass index (-0.8 vs. -0.3 kg/m(2)), systolic blood pressure (-6.6 vs. -2.4 mmHg), total cholesterol (-2.2 vs. +1.5 mg/dl), and the total risk scores (-0.5 vs. -0.1 points). The 15-month intervention program effectively improves and maintains lifestyle and cardiovascular risk, with greater effects of the long-term intervention in females.

High attendance at a lifestyle intervention program is important to reduce risks related to metabolic syndrome in middle-aged Japanese.

Evaluating attendance at health education programs is important to obtain a more comprehensive evaluation of the program impact. This study investigated whether attendance at a lifestyle intervention program in a community setting would reduce risks related to metabolic syndrome. Of 545 subjects with risks related to metabolic syndrome, i.e. overweight, hypertension, dyslipidemia or diabetes, participated in this non-randomized control study, 389 subjects aged 40-71 years completed the surveys at baseline and 27 months. Intervention group (39 males and 168 females) was provided 3 individual counseling plus 28 group sessions conducted monthly on average, whereas control group (64 males and 118 females) received only 7 health information newsletters by mail. Intervention group was subcategorized into two groups according to the median attendance (87.1%): 106 subjects with high attendance (93.8 +/- 4.7%) and 101 subjects with low attendance (68.6 +/- 16.0%). Logistic regression analyses adjusted for age and baseline value showed that among males, the proportion with dyslipidemia risk was lower only in high attendance group compared with that in control group at 27 months [Odds ratio (OR): 0.11 (95%CI 0.02 - 0.51)] and among females, the proportion of overweight was lower only in high attendance group [OR: 0.24 (95%CI 0.07 - 0.81)]. In females, the mean total risk score calculated by adding the number of the 4 risks present decreased only in high attendance group (p < 0.001). In conclusion, high attendance at a lifestyle intervention program impacts the reduction of risks related to metabolic syndrome in a Japanese community setting.

Characterization of Th1 activation by Bartonella henselae stimulation in BALB/c mice: Inhibitory activities of interleukin-10 for the production of interferon-gamma in spleen cells.

This study was conducted to analyze cytokine production mechanisms in mice after Bartonella henselae stimulation. BALB/c mice were inoculated intraperitoneally with 3 x 10(6) colony forming units of B. henselae (Houston-1 strain) twice at 10-day interval. Spleen cells were harvested from the mice and stimulated with the organisms. Following the stimulation, interferon-gamma (IFN-gamma) and interleukin-4 (IL-4), IL-10, IL-12 and tumor necrosis factor-alpha (TNF-alpha) were measured in the culture supernatants of the spleen cells by ELISA. The spleen cells specifically secreted IFN-gamma, but not IL-4, indicating that T helper 1 (Th1) cells were activated following B. henselae stimulation. In addition, IL-10 and TNF-alpha productions were also detected in the culture supernatants of spleen cells. Neutralization of IL-10 in the culture supernatants significantly enhanced the production of IFN-gamma from the spleen cells stimulated with B. henselae. These results indicate that B. henselae predominantly stimulated Th1 cells and resulted in secreting IFN-gamma, however the production was partially inhibited by IL-10, which was produced simultaneously.

Low income, unemployment, and suicide mortality rates for middle-age persons in Japan.

The relationships between age-specific suicide mortality rates and social life factors for all 47 Japanese prefectures in 1980, 1985, and 1990 were assessed by multiple regression analysis after factor analysis on 20 social life indicators. During this period, Japan experienced a secondary oil crisis in 1980-1983 and a bubble economy in 1986-1990. It was concluded that (1) low income was the major determinant which positively affected suicide mortality rate in middle-aged men during a previous 20-yr. period (1970-1990), (2) urbanization was negatively associated with male suicide mortality rates in most of the age classes in the 1980s, (3) unemployment was one of the major determinants of increased suicide mortality rate in middle-age men in the 1980s, and (4) unemployment was the major factor which was inversely associated with suicide mortality rate for elderly women from 1980 to 1990 in Japan.

Tobacco and alcohol tax relationships with suicide in Switzerland.

Previous research has shown an empirical link between tobacco and alcohol use and suicide. If tobacco and alcohol use contribute to suicidal behaviors, then policies designed to reduce the tobacco and alcohol consumption may succeed in reducing suicides as well. To test this hypothesis, correlations for suicide rates with alcohol consumption, taxes on alcohol and tobacco in Switzerland were examined using sets of time-series data from Switzerland in 1965-1994. The tax on tobacco correlated significantly negatively with male standardized suicide rate. The tax on alcohol also correlated significantly with male standardized suicide rate in an autoregressive model. On the other hand, significant relationships with female suicide rate were not found. Policies designed to reduce tobacco consumption are consistent with a benefit of reducing suicides, particularly for men in this sample.

Trends and monthly variations in the historical record of suicide in Japan from 1976 to 1994.

Monthly suicide rates in Japan were analyzed for the period from 1976 to 1994 to clarify trends and recurring effects. The data were separated by sex, and the least-squares method was used. The major findings were (1) a significant positive correlation between unemployment rate and suicide rate for both sexes, (2) the suicide rate was highest in April for both sexes, and (3) an upsurge in male suicide mortality was noted from 1983 to 1990. These findings may well be associated with socioeconomic factors as well as neurobehavioral variables.

Enterprise failures correlate positively with suicide rate for both sexes in Japan.

Correlations among suicide rate, unemployment rate, and enterprise failure were examined from 1976 to 1994 by using a Loess smoothing method. Significant positive correlations were found between suicide rate in men and unemployment and between suicide rate in both sexes and enterprise failure (p < .001).

Stem cell-like differentiation potentials of endometrial side population cells as revealed by a newly developed in vivo endometrial stem cell assay.

BACKGROUND: Endometrial stem/progenitor cells contribute to the cyclical regeneration of human endometrium throughout a woman's reproductive life. Although the candidate cell populations have been extensively studied, no consensus exists regarding which endometrial population represents the stem/progenitor cell fraction in terms of in vivo stem cell activity. We have previously reported that human endometrial side population cells (ESP), but not endometrial main population cells (EMP), exhibit stem cell-like properties, including in vivo reconstitution of endometrium-like tissues when xenotransplanted into immunodeficient mice. The reconstitution efficiency, however, was low presumably because ESP cells alone could not provide a sufficient microenvironment (niche) to support their stem cell activity. The objective of this study was to establish a novel in vivo endometrial stem cell assay employing cell tracking and tissue reconstitution systems and to examine the stem cell properties of ESP through use of this assay. METHODOLOGY/PRINCIPAL FINDINGS: ESP and EMP cells isolated from whole endometrial cells were infected with lentivirus to express tandem Tomato (TdTom), a red fluorescent protein. They were mixed with unlabeled whole endometrial cells and then transplanted under the kidney capsule of ovariectomized immunodeficient mice. These mice were treated with estradiol and progesterone for eight weeks and nephrectomized. All of the grafts reconstituted endometrium-like tissues under the kidney capsules. Immunofluorescence revealed that TdTom-positive cells were significantly more abundant in the glandular, stromal, and endothelial cells of the reconstituted endometrium in mice transplanted with TdTom-labeled ESP cells than those with TdTom-labeled EMP cells. CONCLUSIONS/SIGNIFICANCE: We have established a novel in vivo endometrial stem cell assay in which multi-potential differentiation can be identified through cell tracking during in vivo endometrial tissue reconstitution. Using this assay, we demonstrated that ESP cells differentiated into multiple endometrial lineages in the niche provided by whole endometrial cells, indicating that ESP cells are genuine endometrial stem/progenitor cells.

Suicide mortality of young, middle-aged and elderly males and females in Japan for the years 1953-96: time series analysis for the effects of unemployment, female labour force, young and aged population, primary industry and population density.

Effects of nine social life indicators on age-adjusted and age-specific annual suicide mortality of male and female Japanese population in the years 1953-96 were investigated by multiple regression analysis on time series data. Unemployment rate was significantly related to the age-adjusted mortality in both males and females. Also, female labour force participation was positively related to the male mortality; persons and 65 and above was inversely related to the male mortality. Results on the age-specific mortality indicated that: during the 44 yr, (1) unemployment significantly related with the mortality of young, middle-aged and elderly males and young females; (2) female labour force participation significantly related with the mortality of young and elderly males and young females; aged population significantly related with the mortality of middle-aged and elderly males; (4) young population significantly related with the mortality of young and middle-aged males and females; (5) divorce significantly related with the mortality of middle-aged and elderly males and young males and females; (6) persons employed in primary industries significantly related with the mortality in middle-aged males and young males and females; and (7) population density significantly related with the mortality of middle-aged males and young females.

Lnk negatively regulates self-renewal of hematopoietic stem cells by modifying thrombopoietin-mediated signal transduction.

One of the central tasks of stem cell biology is to understand the molecular mechanisms that control self-renewal in stem cells. Several cytokines are implicated as crucial regulators of hematopoietic stem cells (HSCs), but little is known about intracellular signaling for HSC self-renewal. To address this issue, we attempted to clarify how self-renewal potential is enhanced in HSCs without the adaptor molecule Lnk, as in Lnk-deficient mice HSCs are expanded in number >10-fold because of their increased self-renewal potential. We show that Lnk negatively regulates self-renewal of HSCs by modifying thrombopoietin (TPO)-mediated signal transduction. Single-cell cultures showed that Lnk-deficient HSCs are hypersensitive to TPO. Competitive repopulation revealed that long-term repopulating activity increases in Lnk-deficient HSCs, but not in WT HSCs, when these cells are cultured in the presence of TPO with or without stem cell factor. Single-cell transplantation of each of the paired daughter cells indicated that a combination of stem cell factor and TPO efficiently induces symmetrical self-renewal division in Lnk-deficient HSCs but not in WT HSCs. Newly developed single-cell immunostaining demonstrated significant enhancement of both p38 MAPK inactivation and STAT5 and Akt activation in Lnk-deficient HSCs after stimulation with TPO. Our results suggest that a balance in positive and negative signals downstream from the TPO signal plays a role in the regulation of the probability of self-renewal in HSCs. In general, likewise, the fate of stem cells may be determined by combinational changes in multiple signal transduction pathways.

Structures and physiological roles of 13 integral lipids of bovine heart cytochrome c oxidase.

All 13 lipids, including two cardiolipins, one phosphatidylcholine, three phosphatidylethanolamines, four phosphatidylglycerols and three triglycerides, were identified in a crystalline bovine heart cytochrome c oxidase (CcO) preparation. The chain lengths and unsaturated bond positions of the fatty acid moieties determined by mass spectrometry suggest that each lipid head group identifies its specific binding site within CcOs. The X-ray structure demonstrates that the flexibility of the fatty acid tails facilitates their effective space-filling functions and that the four phospholipids stabilize the CcO dimer. Binding of dicyclohexylcarbodiimide to the O(2) transfer pathway of CcO causes two palmitate tails of phosphatidylglycerols to block the pathway, suggesting that the palmitates control the O(2) transfer process.The phosphatidylglycerol with vaccenate (cis-Delta(11)-octadecenoate) was found in CcOs of bovine and Paracoccus denitrificans, the ancestor of mitochondrion, indicating that the vaccenate is conserved in bovine CcO in spite of the abundance of oleate (cis-Delta(9)-octadecenoate). The X-ray structure indicates that the protein moiety selects cis-vaccenate near the O(2) transfer pathway against trans-vaccenate. These results suggest that vaccenate plays a critical role in the O(2) transfer mechanism.

Lack of association between the IL13 variant Arg110Gln and susceptibility to cedar pollinosis in a Japanese population.

BACKGROUND: Interleukin (IL)-13 has come to be appreciated as a molecule critically involved in allergic inflammatory responses. Recent studies revealed that a common variant in the coding region of the IL13 gene, Arg110Gln, has been implicated in the development of asthma and atopy. METHODS: To assess whether the IL13 variant Arg110Gln is associated with cedar pollinosis, one of the most common atopic diseases in the Japanese population, we examined the Arg110Gln variant using PCR-RFLP to compare the genotype and allele frequencies between 95 patients with cedar pollinosis and 95 healthy control subjects. Relationships between the Arg110Gln variant and the pollinosis-related traits, e.g. rhinitis severity, eosinophil counts in nasal secretion and serum total and allergen-specific IgE levels, were also investigated. RESULTS: The frequencies of the minor allele Gln110 were 25.8% in patients with cedar pollinosis and 30.9% in healthy control subjects (p > 0.05). There was also no significant difference in the genotype frequencies between cases and controls (p > 0.05). In addition, we found no significant association of the Arg110Gln variant with any of the pollinosis-related phenotypes (p > 0.05). CONCLUSIONS: Our data suggest lack of evidence for identifying the variant Arg110Glnat the IL13 locus as a genetic risk factor involved in the development of Japanese cedar pollinosis.

Molecular characterization of a novel nuclear transglutaminase that is expressed during starfish embryogenesis.

We report the constitution and molecular characterization of a novel transglutaminase (EC 2.3.2.13) that starts to accumulate specifically in the nucleus in the starfish (Asterina pectinifera) embryo after progression through the early blastula stage. The cDNA for the nuclear transglutaminase was cloned and the cDNA-deduced sequence defines a single open reading frame encoding a protein with 737 amino acids and a predicted molecular mass of 83 kDa. A comparison of this transglutaminase with other members of the gene family revealed an overall sequence identity of 33-41%. A special sequence feature of this transglutaminase, which is not found in other transglutaminases, is the presence of nuclear localization signal-like sequences in the N-terminal region. Microinjection of hybrid constructs that encode the N-terminal segment fused to reporter proteins into the germinal vesicle of an oocyte produced chimeric proteins by transcription-coupled translation. It was found that the N-terminal segment alone was sufficient to effect nuclear accumulation of an otherwise cytoplasmic protein. These results suggest that the nuclear accumulation of the transglutaminase may play an important role in nuclear remodeling during early starfish embryogenesis.

In vivo cross-linking of nucleosomal histones catalyzed by nuclear transglutaminase in starfish sperm and its induction by egg jelly triggering the acrosome reaction.

A histone heterodimer, designated as p28, which contains an Nepsilon(gamma-glutamyl)lysine cross-link between Gln9 of histone H2B and Lys5 or Lys12 of histone H4, is present in starfish (Asterina pectinifera) sperm. Treatment of sperm nuclei with micrococcal nuclease produced soluble chromatin, which was size-fractionated by sucrose-gradient centrifugation to give p28-containing oligonucleosome and p28-free mononucleosome fractions, indicating that the cross-link is internucleosomal. When sperm nuclei were incubated with monodansylcadaverine, a fluorescent amine, in the presence or absence of Ca(2+), histone H2B was modified only in the presence of Ca(2+). Gln9, in the N-terminal region, was modified, but the other Gln residues located in the internal region were not, suggesting that the modification takes place on the surface of the nucleosome core by the in situ action of a Ca(2+)-dependent nuclear transglutaminase. Treatment of sperm with the egg jelly, which activates Ca(2+) influx to induce the acrosome reaction, resulted in a significant elevation of the p28 content in the nucleus. This is the first demonstration of an in vivo activation of transglutaminase leading to the formation of a cross-link in intracellular proteins.